Share The Seeds
Gardening Area => Advanced Cultivation Techniques => Topic started by: fairdinkumseeds on December 02, 2014, 12:30:07 AM
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Eustrephus latifolius or wombat berry is a cool plant and I reckon it has lots of potential as a mainstream supermarket vegetable, BUT it is still too small and fiddly to clean and sort.
Me using selection to improve size is proving ineffective and taking forever!
SO, I want to make this fella grow bigger roots.
The tubers it makes are great, but quite small @ 2-7cm.
I will have a couple thousand seeds soon. (If I put some real work in I can collect ~10,000 max this season)
What do you recommend I treat the seeds with to encourage potentially beneficial mutations.
Must be reasonably cheap, easy and available. Safe would be cool too...
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http://thenerdygardener.hubpages.com/hub/Australian-Native-Plant-Profile-Wombat-Berry-Eustrephus-latifolius
Bit of info.
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I hope I can be of some help in technical nature, rather than in botanical ways, since I never tried this.
When treating with radiation best results could be archieved with gamma rays, but technically they are complicated to shield (handle) so I would not recommend. X-rays can easily be produced electrically and shielded with reasonable effort, they are pretty much DIY thats why they were discovered and reproduced so long ago. They have proved to be easily capable to change crops characteristics. I can not advise on how much mortality one wants to aim at to have sufficient changes in the viable seeds (and basically less effort growing and selecting). UV Rays will sadly not penetrate deep enough.
(I think I should mention heat is also known to manipulate genes at random but the line between fatal and changed characteristic is very narrow.
The other option I'd consider are chemical treatments, the number of chemical compounds capable of making theese changes (some to frigthening extend, very fast and efficient) is virtually unlimited but many are restricted and one could argue this is within reason. When trying to cause mutation chemically one should take the way of action into consideration, some chemicals for example are faulty build into the DNA during replication (wich doesnt really happen in seeds so its not applicable). Others can recommect the whole "DNA grid" and dont need replication.
Lastly Viruses should be mentioned, they can be build into DNA but I would really not recommend them, let alone know who would quickly hand the right one over to anyone.
Id say considerably safe and obtainable are Nitrosamines and ordinary petrochemical products (in both cases the Vapours should do). Exposure will sure be to be determined in experiments. Smoke will possibly contain benzopyrene amongst other nasty stuff, cold smoking seeds might help. Hydrogen Peroxide in higher doses should do. Benzene and Formaldehyde could do a nice job. Many other chemicals I could think of will possibly be very overkill.
Another idea: in my area and propably many others Colchicum Autumnale can be found. It is known to be "highly toxic" and this is due to a mutagene (1.8% in petals). It works on plants but I cant explain why not on the plant itself. The poison messes with the mitosis. In plants it has been used before to restore fertility in plants that lost the ability to reproduce by crossbreeding. It will create polyploides - again its a narrow area in wich we get viable polyploides, not untreated and not dead seeds. Be aware it will also want to make you polyploide.
Hope you stay safe :) I can advise on producing ionising radiation or nitrosamines.
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Ethyl methanesulfonate
http://www.sigmaaldrich.com/catalog/product/sigma/m0880?lang=en®ion=US
Sodium Azide
http://www.sigmaaldrich.com/catalog/product/sigma/71289?lang=en®ion=US
Chemical mutagens have gained popularity since they are easy to use, do not require any specialised equipment, and can provide a very high mutation frequency. Compared to radiological methods, chemical mutagens tend to cause single base-pair (bp) changes, or single-nucleotide polymorphisms (SNPs) as they are more commonly referred to, rather than deletions and translocations. Of the chemical mutagens, EMS (ethyl methanesulfonate) is today the most widely used. EMS selectively alkylates guanine bases causing the DNA-polymerase to favor placing a thymine residue over a cytosine residue opposite to the O-6-ethyl guanine during DNA replication, which results in a random point mutation. A majority of the changes (70–99%) in EMS-mutated populations are GC to AT basepair transitions [18, 19]. Mutations in coding regions can be silent, missense or nonsense. In noncoding regions, mutations can change promoter sequences or other regulatory regions, resulting in up- or downregulation of gene transcription. Aberrant splicing of mRNA, altered mRNA stability and changes in protein translation may also occur as a result of mutagenesis.
Other mutagens such as sodium azide (Az) and methylnitrosourea (MNU) are also used and often combined into an Az-MNU solution. Genetically, Az-MNU predominantly causes GC to AT shifts, or AT to GC shifts. Thus, contrary to EMS, a shift can happen in either direction [18]. All three chemical mutagens are, as can be expected, strongly carcinogenic and should be handled with extreme care. Unlike EMS, MNU is both sensitive to shock and unstable above 20°C making it complicated to work with. In contrast to EMS and MNU, which are both liquid, Az is a solid dust in its ground state and the additional step of first dissolving the acutely toxic and volatile substance before application makes it less attractive to handle.
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I found something cute :) I found many Colchicum recently that flower at the "wrong" time which is said to be rare but becoming more often recently. The big lot of Colchicum was in a public park :O with children going all excited about them ... :O
Also I found a look alike that one would not want to induce Polyploidy, simply because it doesnt :D Note the leaves, true Colchicum have different leaves and will never show leaves and flower at the same time.
Yes they can go all the way from white to nearly blue and everything in between, also theres many different Colchicum with Autumnale being most present in my area, all will unduce polyploidy when something is treated with them or someone handles them withour care.
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Wow, in a park with kids! There should be a warning sign. Very pretty flowers though.
Still experimenting on cannabiseed, soaked in 0.05% Oryzalin, solution for three days.
Been meaning to take some pics and make an update. it didn't seem to improve them.
Most died, some sprouted and died immediately, and some died later, some survivors.
More details in that thread soon w/pics. The natural Colchicum seems most promising.
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Colchicum Autumnale
Keep an eye out for seeds. I planted heaps a while back but no luck. Might have been duds or maybe its not long enough, but all the same would love a couple fresh ones.
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Lets give thit a lik bump!
I have been out checking and guess what I found, right, lots of Colchicum. I guess on average I have ~5 plants every square meter in the Colchicum place :D Every plant produces one or two pods that'll contain the seeds. I snapped some photos of the plants showing first signs of that pod and sliced a pod open to get a idea how mature the seeds are and how many they'll be per plant. I know, I need to clean out this camera :D
Its always after doing so that I start to wonder, is it safe to handle ? Last time we talked Colchicum I considered gloves but yeah :D
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About how much colchicine are we talking here btw ? I'm set getting those seeds, but regarding seed or plant treatment, I am considering using gout medication, pills with 0.6mg Colchicine, pure and less risk of messing up terribly.
So what concentrations are we looking for when we want to treat either seeds or plants ?
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So still somewhat teased by the idea, maybe less the result more all the fun involved I stick my nose into ionising radiation once in the while and wonder how everyone can make some at home (LOL) without too much effort or even going illegal or something (might want to note that in most countries there is laws about sending certain frequencies at certain transmission power without shielding). So I still consider X-rays affordable, easy, controllable, relatively safe and so on and I finally found a way how people get around producing or acquiring the X-Ray tube.
Simply: Use an old electric tube amplifier and overload that, I have looked up several that are still in production or on stock, common, cheap and will operate under conditions that have never been the intention, namely: DY802 DY86 EY87 GY501 1Z21P 1Z11pP GP5, they wont produce mad X-ray as your doctors will but honestly, you and your neighbors dont want that, yet they will make decent radiation. Seeds or plants sitting in a suffieciently shielded and maybe cooled chamber should mutate pretty happily. Calculation for the DY802 ( ~1$) shows that you should be able to dump 2 Watts in it at 50.000 Volts, the rule of thumbs say that 1 - 2% will be transformed to radiation. You should be able to easily keep yourself safe from that if you dont take it to bed.
I think if I ever touch the subject I will cover this route, and obvious the Colchicum since its right at my place. But also note: both things are not to be compared, completely different.
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I'm currently experimenting with Oryzalin sold as Surflan. You should use young seedlings rather than seeds as its an herbicide. In some cases its more efficient than Colchicum. Oryzalin is extremely cheap due to the working concentration.
Another cheap mutagen which is safe is caffeine. I didn't find the mg used but should be easy to test. From what i remember you apply it to soaked seeds or freshly germinated seeds. Not very effective but does work.
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You haven´t tried heterosis yet?
Mycorrhiza could help the plant to get nutrients more efficiently and eventually protect from parasites or diseases locally in or on the root-system.
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But we want Muuuuutants :D like all kind of scary birthdefects until we find one adorable
Gigantism, Miniatism, Colors, shapes ...
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Another cheap mutagen which is safe is caffeine. I didn't find the mg used but should be easy to test. From what i remember you apply it to soaked seeds or freshly germinated seeds. Not very effective but does work.
So a seed soak in coffee instead of water could work??? I'm not sure how safe I can say caffeine is... pure caffeine can be quite toxic, the FDA hasn't outlawed buying it in US but has put warnings against people buying due to people accidentally consuming too much... although we're talking about use as a plant mutagen and I'm uncertain how dangerous it is with regard to absorption through skin/inhalation.
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I think you need better concentration as in coffee, obviously you could make strong coffee but all the other stuff in there ? Well luckily caffeine s|_|bl1mes which mean the Xtr4cti0n is as easy as it can get (please delete if rules no no i modified keywords to not turn out on any search) youd just want to heat it in a metal pot that has good thermal conductivity all around, flip a glass lid around, put it on and fill the cavity with ice. And use it powdered dont try to make coffee, reduce it and sublime it the residue will be from out of this world.
Pure caffeine tastes yuck yuck yuck so please dont try :D
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There are caffeine pills for sale. Jet-Alert is 200 mg per pill so easy to experiment with known amounts. I will need to find the published article but I'm sure they presoaked the seeds rather than seedlings. I'd say 1 mg per mL would be an easy starting point and you can test higher and lower concentrations.
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Starting a trial with 200mg caffeine pills. Seed soak in hundreds of cups, 1/4pill, 1/2pill, 3/4pill, etc etc etc.
Tried surflan, but turns out its a very effective weedkiller on wombat berry, even at really low doses....
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You should presoak seeds in water for a few hours or days depending on the seeds used and germination speed. Then soak in caffeine for 24 hours followed by 24 hours water soak to remove excess caffeine on seed coat. The concentration of caffeine was quite low so you should dialate greatly. 200 mg would go a long way according to this published article. May still be worth testing higher concentrations though it seems less is more.
http://omicsonline.com/open-access/studies-on-the-effect-of-caffeine-on-growth-and-yield-parameters-in-helianthus-annuus-l-variety-modern-0974-8369-1-014.pdf
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FDK, whats your design for evaluation, do you have control groups ? Will you keep track of germination rates so we do have data about how coffeine affects germination, if it has done something will then be the later question :)
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Was just gonna put a heap of disposable cups on a table, 20seeds per cup. Various concentrations caffeine from zero(control) to heaps, various time soaks, 1 hr, 6hrs, 12hrs, 24hrs, 2days, 4days.
Rinse before planting, cross fingers, see what happens.
Surviving sprouts get photographed daily/weekly/monthly.
3months later tubers as big as footballs, instead of peanuts.
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Never heard or read about caffeine being used as a mutagen but if its a dna targeted displacement reaction a ph buffer helps the chemical react with the dna strands while lowering the liklyhood of seed death from chemical shock - i dont believe this is needed when doing rna mutations
Phosphoric acid is probly the best way to achieve this from my understanding
oh and i believe i remember reading that citric acid is a popular buffer in labs aswell
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Me too, but google shows theres several accounts on coffeine being mutagenic in funghi bacteria plants amd animals.
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Very interesting - ill have to look into that
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This might have similar effect like Colchicine: https://en.m.wikipedia.org/wiki/Chelidonine
But thats just a guess.
However I got a rather small amount of seeds now :)
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Question about inducing polypoloidity. Wouldent it b more effective to give small doses of a mutagen to a plant over time and harvest the seed. It seems like a fully established plant could take higher doses of the mutagen over a longer period, increasing your chances. You could even treat several species in one genus and cross them. This should work. And could I make a polypoloidal hybiscus for instance, into a tatrapoloid? How would I know?
I was just reading https://en.m.wikipedia.org/wiki/Triangle_of_U so cool.
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Question about inducing polypoloidity. Wouldent it b more effective to give small doses of a mutagen to a plant over time and harvest the seed. It seems like a fully established plant could take higher doses of the mutagen over a longer period, increasing your chances. You could even treat several species in one genus and cross them. This should work. And could I make a polypoloidal hybiscus for instance, into a tatrapoloid? How would I know?
I was just reading https://en.m.wikipedia.org/wiki/Triangle_of_U so cool.
I think youd get a few lightly mutated genomes with a high chance of all kill switches still functioning and all, the plant will cut those of and not let their genes anywhere near their inheritance line - your seeds will probably be fine.
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Interesting. Hence the seed soaking being the most effective option. I have many seeds and would like to do an experiment soon.
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Im not 100% sure of your question but if you try to cause a dna mutation in a living plant you would have a lower success rate because the plant has a dna repairing mechanism but would also have a lower fatality rate
If your shooting for mutations its generally better to mutate the seeds of a plant because they are not living yet and have no way to repair the dna you damage with the mutagen, although this has a high fatality rate for some seed species so you would wanna start with a lot of them to offset this
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Another debatable question. If I get a successful mutation in a vegitable, is it safe to,consume?
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That question has been debated for decades - but the way i personally look at it is.. just about everything grown in this country is geneticaly modified to produce higher yields for the farmer, knowingly or not ive been eating them my whole life and it hasnt killed me, harmed me, or had adverse effects, i have never met someone face to face who is ill or dying because the have eaten canned corn or green beans so im willing to bet that gmo plants are safe to consume
But theres thousands of pages of documents on both sides that says the other is wrong
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A new plant is a new plant and usually has predictable but might have supeising new features... especially in the case of random (!) mutation you might be suprised by the new features. The odds are, that if the parent plant was edible the new alternated is too, but every new plant has to be investigated to be sure, no matter if it was crossbred GMd or mutated.
The more interesting question is: Would the change in the plant affect the experience of eating it in any way, and even more: Would it affect it in a positive way. Imagine the number of possible favorable changes and overall possibly changes (which are near endless) it becomes obvious: Your odds of having improved something the way you wanted it are low.
Personally I think a real-deal GMO is even more safe than the original plant because all the fear and controversy got us into imposing laws on the inspection of GMOs before release that have so high requirements, they might even be finding things that are in the original and modified organism that we didn't know about yet.
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+1 for that 3rd paragraph bubble... very well said